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Tatiana Romanova is introduced as a corporal in Soviet Army Intelligence, assigned to work in the Soviet Embassy in Istanbul as a cipher clerk. Her superiors, in connection with SMERSH, plan to sow dissent in the intelligence community by murdering and discrediting a significant figure in western intelligence. This was to be achieved by the murder of James Bond.

Her commanding officer is Rosa Klebb — in the screenplay adaptation, she is secretly an agent for SPECTRE — who manipulates her into believing that she is on an important mission for her country, when she is in fact merely a pawn in the terrorist organization's latest bid to destroy the British Secret Service.

In the novel, Klebb is actually a member of the Soviet government, commander in chief of the Otdyel II section of the SMERSH. Thus, Romanova works for the Soviets, who assign her the mission of seducing Bond and having him take her to England to deliver a SPEKTOR code machine, as well as plant false information, before being rescued from a political prison and returned to Russia.

Once in Istanbul, she contacts Darko Kerim Bey (Ali Kerim Bey in the film) and tells him her plans: she would willingly defect from the Soviet Union and take with her the precious LEKTOR/SPEKTOR only if Bond assisted the operation. She claimed that she had fallen in love with the man from a picture she had seen in a secret file and wanted to live with him.

Both M and Bond believe this to be a trap, but the prize is valuable enough to go for it. They react just as the main instigator of the plot, chess champion Kronsteen, had predicted. Bond then flies to Istanbul and contacts Kerim, spending several days there, waiting for contact. After a riot at a Gypsy camp, Bond returns to his hotel room and finds Romanova walking across the room and getting into his bed, wearing only a black velvet choker and black stockings. They make love, but are secretly filmed by Klebb's minions via a one-way mirror. This tape is explicitly planned to be used to embarrass MI6.

In the film, after meeting her again to verify the authenticity of her information, they blow up the Soviet Embassy in Istanbul to cover their escape. With the help of Kerim, they board the Orient Express and depart for Trieste and the Italian frontier. As a part of Kronsteen's strategy, the SMERSH/SPECTRE assassin Donald 'Red' Grant kills Kerim. Bond contacts Grant, who is pretending to be an agent named Nash. After sedating Romanova, Nash's identity is revealed to Bond, who then fights Grant in their train room. Prior to the fight, Grant explains that he is going to kill Bond and then execute the sleeping Romanova with Bond's gun, making it look like a murder-suicide. Bond finally defeats Grant and takes Romanova to Venice. It is there they meet Klebb again who, in an attempt to retrieve the LEKTOR and kill Bond, disguises herself as a maid and tries to eliminate the agent with a dagger-tipped shoe poisoned with blowfish venom. Romanova shoots Klebb, thus saving Bond. She and Bond are last seen on a boat in Venice, with Bond dropping the incriminating negatives into the canal.

Similarly, in the novel, they board the Orient Express with Kerim, planning to travel to England over the course of four days. Kerim, instead of being killed by Grant, is killed by a Russian agent named Benz who had boarded the train earlier and also is killed in the struggle. This prompts the bombing of the Soviet Embassy in Istanbul in retaliation. Despite this, Bond elects not to leave the train for a plane or the embassy, after having fallen for Romanova and not wanting to cut their time short. Grant pretends to be Nash, an MI5 agent sent by M in response to the death of Kerim. After sedating Romanova, under the guise of standing guard over Bond, he waits until they are both asleep, and plans to murder them. However, due to his vanity, he taunts Bond, revealing details of a meeting with Rosa Klebb. This allows him to disarm and save Romanova's life. It is unclear as to what ultimately becomes of Tatiana in the novel as in her last appearance, she is still heavily affected by the sedatives, sleeping in the British Embassy, while Bond confronts Klebb. It is presumed that she has been arrested and/or released by the British.

The reception of the Tatiana Romanova's character was generally excellent, with many people going to watch the movie and enjoying the scenes featuring her. People were especially fond of the Russian ethnicity and culture, and the thought of a Russian defecting from the motherland to join the west. Especially during the Cold War, her character was used as a symbol of Russian women who wanted to defect Russia and join the west. The thought was that the west had better and more manly men with more money. In addition, the perception was that western men cared about women, as opposed to the perception of Russian men, which mostly relied on alcohol and violence.